使用液相色谱/高分辨质谱法对马中的雄甾烯-1,4,6-三烯-3,17-二酮进行代谢研究。,The Journal of Steroid Biochemistry and Molecular Biology

当前位置： X-MOL 学术 › J. Steroid Biochem. Mol. Biol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

使用液相色谱/高分辨质谱法对马中的雄甾烯-1,4,6-三烯-3,17-二酮进行代谢研究。
The Journal of Steroid Biochemistry and Molecular Biology ( IF 2.7 ) Pub Date : 2015-06-03 , DOI: 10.1016/j.jsbmb.2015.05.011
Wai Him Kwok ₁ , Gary N W Leung ₁ , Terence S M Wan ₁ , Peter Curl ₂ , Peter J Schiff ₂

Affiliation

Androsta-1,4,6-triene-3,17-dione（ATD）是不可逆的甾体芳香酶抑制剂，以补充剂形式销售。据报道可有效减少雌激素的生物合成并显着增加人体内内源性类固醇如二氢睾丸激素和睾丸激素的水平。据报道，人类体育运动中滥用了ATD。已经研究了其在人体内的代谢，并且已经报道了马中ATD的体外代谢研究，但是，关于其在马中的生物转化和消除知之甚少。本文介绍了马中ATD的体内外代谢研究，目的是鉴定具有最长检测时间以控制ATD滥用的目标代谢物。使用均质的马肝进行ATD的体外代谢研究。发现ATD被广泛代谢，并且由于灵敏度不足，其代谢物无法通过气相色谱/质谱（GC / MS）轻松表征。因此，采用液相色谱/高分辨率质谱（LC / HRMS）鉴定体外代谢物。观察到的主要生物转化是在C3或C17位上烯烃基和/或酮基还原的组合。另外，观察到D-环中的单羟基化以及在C17位的烯烃基和/或酮基的还原。十四种体外代谢物，包括雄激素1,4,6-trien-17-ol-3-one（M1a，M1b）和雄激素4,6-diene-3,17-dione（M2）的两种差向异构体（M3），androsta-4,6-diene-17β-ol-3-one（M4），androsta-4,6-diene-3-ol-17-one（M5），双烯酮和Epi-boldenone（M6a，M6b），羟基化的androsta-1,4,6-trien-17-ol-3-one的四个立体异构体（M7a至M7d）和androsta-1,4-diene-16α，17-diol的两个差向异构体（M8a， M8b）。通过使用LC / HRMS与真实的参考标准品进行匹配，可以确认除M1a，M5，M7a至M7d以外的所有代谢物的身份。为了进行体内代谢研究，分别通过胃管向两种纯种胶与800 mg ATD一起给药。在给药后尿液中检测到了ATD，以及十四种体外代谢物中的十二种，包括M1a，M1b，M2，M4，M5，M6，M7a至M7d，M8a和M8b。质谱解释性地确定了另外两种尿代谢产物，即羟基化的雄甾烷-4,6-二烯-17-醇-3-酮的立体异构体（M9a，M9b）。还观察到睾丸激素水平升高。在给药后的血液样本中，仅鉴定了母体药物M1b和M2。这项研究表明，检测尿液中的代谢物M1b（雄激素-1,4,6-三烯-17β-ol-3-one）或M4（均以硫酸盐结合物的形式排泄）是检测ATD的最佳方法，给药后最多可检测77小时。血浆选择的分析物是M1b，在给药后最多28小时即可检测到。

"点击查看英文标题和摘要"

Metabolic study of androsta-1,4,6-triene-3,17-dione in horses using liquid chromatography/high resolution mass spectrometry.

Androsta-1,4,6-triene-3,17-dione (ATD) is an irreversible steroidal aromatase inhibitor and is marketed as a supplement. It has been reported to effectively reduce estrogen biosynthesis and significantly increase the levels of endogenous steroids such as dihydrotestosterone and testosterone in human. ATD abuses have been reported in human sports. Its metabolism in human has been studied, and the in vitro metabolic study of ATD in horses has been reported, however, little is known about its biotransformation and elimination in horses. This paper describes the in vitro and in vivo metabolism studies of ATD in horses, with an objective of identifying the target metabolites with the longest detection time for controlling ATD abuse. In vitro metabolism studies of ATD were performed using homogenized horse liver. ATD was found to be extensively metabolized, and its metabolites could not be easily characterized by gas chromatography/mass spectrometry (GC/MS) due to insufficient sensitivity. Liquid chromatography/high resolution mass spectrometry (LC/HRMS) was therefore employed for the identification of in vitro metabolites. The major biotransformations observed were combinations of reduction of the olefin groups and/or the keto group at either C3 or C17 position. In addition, mono-hydroxylation in the D-ring was observed along with reduction of the olefin groups and/or the keto group at C17 position. Fourteen in vitro metabolites, including two epimers of androsta-1,4,6-trien-17-ol-3-one (M1a, M1b), androsta-4,6-diene-3,17-dione (M2), boldione (M3), androsta-4,6-diene-17β-ol-3-one (M4), androsta-4,6-diene-3-ol-17-one (M5), boldenone and epi-boldenone (M6a, M6b), four stereoisomers of hydroxylated androsta-1,4,6-trien-17-ol-3-one (M7a to M7d), and two epimers of androsta-1,4-diene-16α,17-diol (M8a, M8b), were identified. The identities of all metabolites, except M1a, M5, M7a to M7d, were confirmed by matching with authentic reference standards using LC/HRMS. For the in vivo metabolism studies, two thoroughbred geldings were each administered with 800 mg of ATD by stomach tubing. ATD, and twelve out of the fourteen in vitro metabolites, including M1a, M1b, M2, M4, M5, M6, M7a to M7d, M8a and M8b, were detected in post-administration urine. Two additional urinary metabolites, namely stereoisomers of hydroxylated androsta-4,6-dien-17-ol-3-one (M9a, M9b), were tentatively identified by mass spectral interpretation. Elevated level of testosterone was also observed. In post-administration blood samples, only the parent drug, M1b and M2 were identified. This study showed that the detection of ATD administration would be best achieved by either monitoring the metabolites M1b (androsta-1,4,6-trien-17β-ol-3-one) or M4 (both excreted as sulfate conjugates) in urine, which could be detected for up to a maximum of 77 h post-administration. The analyte of choice for plasma is M1b, which could be detected for up to 28 h post administration.

更新日期：2019-11-01

点击分享查看原文

点击收藏

阅读更多本刊新发论文